RRC ID 50101
著者 Peek J, Harvey C, Gray D, Rosenberg D, Kolla L, Levy-Myers R, Yin R, McMurry JL, Kerscher O.
タイトル SUMO targeting of a stress-tolerant Ulp1 SUMO protease.
ジャーナル PLoS One
Abstract SUMO proteases of the SENP/Ulp family are master regulators of both sumoylation and desumoylation and regulate SUMO homeostasis in eukaryotic cells. SUMO conjugates rapidly increase in response to cellular stress, including nutrient starvation, hypoxia, osmotic stress, DNA damage, heat shock, and other proteotoxic stressors. Nevertheless, little is known about the regulation and targeting of SUMO proteases during stress. To this end we have undertaken a detailed comparison of the SUMO-binding activity of the budding yeast protein Ulp1 (ScUlp1) and its ortholog in the thermotolerant yeast Kluyveromyces marxianus, KmUlp1. We find that the catalytic UD domains of both ScUlp1 and KmUlp1 show a high degree of sequence conservation, complement a ulp1Δ mutant in vivo, and process a SUMO precursor in vitro. Next, to compare the SUMO-trapping features of both SUMO proteases we produced catalytically inactive recombinant fragments of the UD domains of ScUlp1 and KmUlp1, termed ScUTAG and KmUTAG respectively. Both ScUTAG and KmUTAG were able to efficiently bind a variety of purified SUMO isoforms and bound immobilized SUMO1 with nanomolar affinity. However, KmUTAG showed a greatly enhanced ability to bind SUMO and SUMO-modified proteins in the presence of oxidative, temperature and other stressors that induce protein misfolding. We also investigated whether a SUMO-interacting motif (SIM) in the UD domain of KmULP1 that is not conserved in ScUlp1 may contribute to the SUMO-binding properties of KmUTAG. In summary, our data reveal important details about how SUMO proteases target and bind their sumoylated substrates, especially under stress conditions. We also show that the robust pan-SUMO binding features of KmUTAG can be exploited to detect and study SUMO-modified proteins in cell culture systems.
巻・号 13(1)
ページ e0191391
公開日 2018-1-19
DOI 10.1371/journal.pone.0191391
PII PONE-D-17-00761
PMID 29351565
PMC PMC5774762
MeSH Amino Acid Sequence Catalytic Domain / genetics Conserved Sequence Cysteine Endopeptidases / chemistry Cysteine Endopeptidases / genetics Cysteine Endopeptidases / metabolism* Fungal Proteins / chemistry Fungal Proteins / genetics Fungal Proteins / metabolism* Genetic Complementation Test Kluyveromyces / genetics Kluyveromyces / metabolism Models, Molecular Mutant Proteins / chemistry Mutant Proteins / genetics Mutant Proteins / metabolism Protein Binding Protein Processing, Post-Translational Saccharomyces cerevisiae / genetics Saccharomyces cerevisiae / metabolism Saccharomyces cerevisiae Proteins / chemistry Saccharomyces cerevisiae Proteins / genetics Saccharomyces cerevisiae Proteins / metabolism* Sequence Homology, Amino Acid Small Ubiquitin-Related Modifier Proteins / metabolism* Stress, Physiological Sumoylation Temperature
IF 2.74
引用数 3
リソース情報
酵母 K. maxianus BY28356